Figure 3

CSM regulates expression of TLR4 via ROS. (A) MDMs (5 × 10⁶ cells) were stimulated by CSM (0.03, 0.06 and 0.12 OD) for 4 h with and without pretreatment with NAC (10 mM) for 30 min. RNA was extracted and TLR4 and GAPDH gene expression were quantified by real-time PCR. Results are expressed as copies of TLR4 vs. copies of GAPDH gene. (B) MDMs were preincubated with naturalizing anti-TLR4 or isotype control antibodies for 30 min and then stimulated with CSM (0.06 OD) for 4 h or LPS (100 ng/ml) and mRNA levels of TLR4 was determined by real-time PCR method. Values are expressed as mean +/- S.E.M (n = 3).*P < 0.05,***p = 0.001 significantly different compared to control and # P < 0.05 significantly different compared to CSM stimulated (n = 3).